巨细胞病毒及相关miRNAs在高血压发病中作用和机制研究

Human cytomegalovirus (HCMV) is a double stranded DNA virus belonging to a member of the beta-herpesvirus group. The results from the population association study have been demonstrated that HCMV and a hcmv-miR-UL112, one of the HCMV-encoded microRNAs(miRNAs) were associated with essential hypertension (EH). However, the underlying pathologic mechanisms are poorly understood. And cytomegalovirus (CMV) encodes at least 21 mature miRNAs. In addition to expressing their own miRNAs, CMV infections can result in changes in the expression of the host cellular miRNAs that benefit virus replication. Our preliminary experiments have shown that six HCMV related miRNAs expressions were changed in hypertensive patients, and associated with HCMV IgG antibody titers and hypertension. Therefore, we hypothesized that in Wistar-Kyoto rat(WKY) of CMV infections, because of CMV-encoded miRNAs and target genes interaction with the host cellular changed miRNAs and target genes, resulting in inflammation response,vascular proliferation and remodeling as well as vascular endothelial dysfunction , then CMV promotes the development of hypertension. To prove this hypothesis, WKY and aortic endothelial cells and smooth muscle cells are infected with mouse cytomegalovirus (MCMV) as the disease model, miRNAs expression microarray, signature miRNAs and corresponding target genes screening and validation as well as miRNAs gene knock-down and overexpression lentiviral vectors are employed. We intend to study the effects of the differentially expressed miRNAs and target genes expression changes on WKY blood pressure ,inflammation response,vascular smooth muscle proliferation and remodeling,left ventricular hypertrophy as well as vascular endothelial structure and function based on vivo, tissue, cell and molecular levels. Eventually, we clarify the effects of CMV on hypertension and its mechanisms underlying miRNAs and to provide a new clinical target for the prevention and treatment of hypertension.

源于人群关联研究显示人巨细胞病毒(HCMV)及HCMV编码一种微小RNA(miRNA) hcmv-miR-UL112与高血压相关,机制不明。巨细胞病毒(CMV)至少编码21种成熟miRNAs,并导致宿主体内miRNAs表达发生改变。预实验发现HCMV相关6个miRNAs在高血压患者表达发生改变，且与HCMV抗体滴度和高血压相关。我们推测：CMV通过CMV编码miRNAs与宿主体内发生改变的miRNAs及靶基因交互作用,导致炎症反应、血管重塑及内皮结构功能障碍,促进了高血压发生。为此本项目以鼠CMV感染WKY及主动脉内皮细胞和平滑肌细胞为疾病模型,采用miRNAs表达谱芯片、差异表达miRNAs及靶基因筛选验证、miRNAs慢病毒载体等方法,从不同层面探讨差异表达miRNAs及靶基因变化对WKY血压、炎症反应及血管增殖重构和心肌肥厚等影响,从miRNAs角度阐明CMV在高血压发病中作用和机制

高血压(EH) 是一种由环境因素和遗传因素及表观遗传等相互作用的多基因遗传性疾病。随着研究进展，人们发现EH的发生与人巨细胞病毒 (HCMV)感染及HCMV编码的 miRNAs变化有关。流行病学调查显示全球范围内HCMV感染率为40-100%，HCMV感染人体后终身存在，并产生持续的潜伏感染。HCMV感染人体后，为满足HCMV复制，可导致宿主细胞内miRNAs表达发生改变，但是否参与EH发生发展上不清楚。本项目以鼠巨细胞病毒（MCMV）感染诱发C57BL/6小鼠高血压为模型，从整体、组织和细胞分子水平探讨了MCMV及MCMV感染诱发的差异表达miRNA在高血压发生发展中的作用和机制，研究证实：（1）MCMV感染诱发小鼠出现高血压的同时，外周血检测到118个差异表达miRNAs，上调91个，下调27个；（2） MCMV感染可引起宿主体内和MCMV自身编码的miRNAs表达均发生改变；（3） MCMV感染可诱导宿主体内与高血压相关的32个miRANs出现了差异表达，上调26个，下调6个；（4）mmu-miR-1929-3p、mcmv-miR-m01-4-5p验证结果与miRNAs-seq一致且mmu-miR-1929-3p与内皮素A型受体（ETAR）的3’-UTR区存在靶向关系；（5）MCMV感染促进高血压和心血管重塑发生发展，其机制与MCMV感染诱导下调的miR-1929-3p通过ETAR活化NLRP3炎症小体有关，miR-1929-3p过表达可以改善MCMV所致心血管不良重塑，其作用机制是通过抑制ETAR的表达从而抑制NLRP3炎症小体激活实现。该研究成果首次发现宿主体内miR-1929-3p表达变化与MCMV感染引起的EH具有潜在关系，通过抗HCMV药物联合miR-1929-3p和靶基因ETAR干预有望成为MCMV感染诱发EH防治的新靶点。